Background: Drug resistant Mycobacterium tuberculosis is one of the serious public healthrntrouble that intimidating progress made in tuberculosis cases and control in several countriesrnincluding Ethiopia. Rifampicin resistance is an indicator for drug-resistant Mycobacteriumrntuberculosis, because it disclose the existence of more than 90% Isoniazid resistance. Earlyrndetection of drug-resistant tuberculosis is crucial for patient management and infection control. rnObjective: This study was designed to assess the prevalence of Mycobacterium tuberculosis, itsrnRifampicin-resistance pattern and associated factors among presumptive Pulmonary and ExtrarnPulmonary Tuberculosis patients at Madda Walabu University, Goba Referral Hospital,rnSoutheast Ethiopia. rnMethods: Hospital based cross-sectional study design was carried out from October 22, 2020 tornFebruary 7, 2021. Detection of Mycobacterium tuberculosis and resistance to Rifampicin patternrnwas determined by using GeneXpert MTB/RIF assay. Data were entered and analyzed by SPSSrnversion 23.0. Bivariate and multivariate analyses were used to examine the relationship betweenrndependent and independent variables. Less than 0.05 P-value was used to show significance. rnResults: A total of 301 presumptive tuberculosis patients were included in the study; of these, 46rn(15.3%) of them were identified as having Mycobacterium tuberculosis by the GeneXpertrnMTB/RIF assay, 2/46 (4.35%) were resistant to Rifampicin and 4/46 (14.8%) patients werernTB/HIV co-infected. From the total of M. tuberculosis detected 40 (16.7%) were identified inrnpulmonary and 6 (9.8%) were in extra-pulmonary presumptive patients. Rifampicin-resistant M.rntuberculosis was detected in 2 patients who had a history of taking Anti-tuberculosis drugs andrnno in new patients. rnConclusion and recommendation: Previous history of tuberculosis treatment and having closerncontact history with tuberculosis patients were found as an important associated factors thatrnenhance the prevalence of tuberculosis. This indicates the mandate to make better and overseernthe treatment protocol to control the burden of tuberculosis.